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目的 :探讨人B细胞淋巴瘤活检组织中肿瘤细胞膜表面免疫球蛋白VH(smIgVH)基因片段能否在动物体内激发特异性抗独特型抗体。方法 :以RT PCR法获得IgVH 基因片段 ,以小鼠单核细胞趋化因子 (MCP 3)基因作为佐剂分子 ,进行重组PCR获得MCP 3和VH 基因片段的融合基因 ,克隆在真核表达载体 pcDNA3.1中 ,构建DNA疫苗质粒 pcDNA/MCP BVH。通过脂质体转染验证该质粒在真核细胞COS 7中的表达。结果 :通过上述方法获得了以活检组织肿瘤细胞mIgVH区基因片段 ;成功地构建了DNA疫苗质粒 pcDNA/MCP BVH。体外瞬时转染实验证明 ,该质粒能够在真核细胞COS 7中正确表达。结论 :成功地构建重组表达质粒 pcDNA/MCPBVH,在体外能够正确表达 ,为进一步研制抗B细胞淋巴瘤基因疫苗奠定了基础
Objective: To investigate whether the immunoglobulin VH (smIgVH) gene fragment on the surface of human cell B-cell lymphoma biopsies can stimulate specific anti-idiotypic antibodies in animals. Methods: The IgVH gene fragment was obtained by RT-PCR and the fusion gene of MCP 3 and VH gene fragment was obtained by using recombinant mouse MCP-3 gene as adjuvant. The fusion gene was cloned into eukaryotic expression vector pcDNA3.1 to construct DNA vaccine plasmid pcDNA / MCP BVH. The plasmid was verified for its expression in eukaryotic COS7 by lipofection. Results: The mIgVH gene fragment was obtained by biopsy. The DNA vaccine plasmid pcDNA / MCP BVH was successfully constructed. Transient transfection in vitro experiments show that the plasmid can be correctly expressed in eukaryotic COS7. Conclusion: The recombinant plasmid pcDNA / MCPBVH was successfully constructed and could be correctly expressed in vitro, which laid the foundation for the further development of anti-B cell lymphoma gene vaccine